An intein with genetically selectable markers provides a new approach to internally label proteins with GFP

BMC Biotechnology

Table 1 Plasmids

Name	Base plasmid	Promoter	Origin/marker	Extein[intein]extein
pGPch-1	pGEX-4T-1	tac	ori/Amp	GST[Pch PRP8]HIS₆
pRR01	pGEX-4T-1	tac	ori/Amp	GST[Pch PRP8 linker]HIS₆
pRR02	pGEX-4T-1	tac	ori/Amp	GST[Pch PRP8 G418^R]HIS₆
pRR03	pGEX-4T-1	tac	ori/Amp	GST[Pch PRP8 HIS5]HIS₆
pRR04	pGEX-4T-1	tac	ori/Amp	GST[Pch PRP8 HygB^R]HIS₆
pRR05	pGEX-4T-1	tac	ori/Amp	GST[Pch PRP8 LexA-VP16]HIS₆
pRR11	pET	T7	ori/Amp	GFP[Pch PRP8 linker]FLAG
pRR12	pET	T7	ori/Amp	GFP[Pch PRP8 G418^R]FLAG
pRR13	pET	T7	ori/Amp	GFP[Pch PRP8 HIS5]FLAG
pRR14	pET	T7	ori/Amp	GFP[Pch PRP8 HygB^R]FLAG
pRR15	pET	T7	ori/Amp	GFP[Pch PRP8 LexA-VP16]FLAG
pRR21	pBM258	Gal	cen/ura3	NLS-GFP[Pch PRP8 linker]FLAG
pRR22	pBM258	Gal	cen/ura3	NLS-GFP[Pch PRP8 G418^R]FLAG
pRR23	pBM258	Gal	cen/ura3	NLS-GFP[Pch PRP8 HIS5]FLAG
pRR24	pBM258	Gal	cen/ura3	NLS-GFP[Pch PRP8 HygB^R]FLAG
pRR25	pBM258	Gal	cen/ura3	NLS-GFP[Pch PRP8 LexA-VP16]FLAG
pRR26	pBM258	Gal	cen/ura3	NLS-GFP (no intein)

All plasmids were constructed in this study except pGPch-1 [19]. In the table the promoter that drives expression of the extein-intein is indicated.

ISSN: 1472-6750